## APPENDIX E. Summary of Aquati c Ecological Effects Data for Endosulfan

This appendix is a summary of the data be ing evaluated for ecologi cal effects on aquatic organisms. Each section consists of severa l sets of data. Tables presenting the data submitted with the application for registrati on are presented first. Within a taxonomic group, the lowest value from this table is used to set a toxicity reference value that is used in evaluating the data from the open liter ature—from EPA’s ECOTOX database. Only the studies associated with the ECOTOX data that have values less than these toxicity reference values are examined further. If any of these latter values are acceptable, then that value becomes the final value used in the risk assessment for that particular taxonomic group.
Because the stability of endosulfan TGAI in a quatic tests is a concern due to its volatility, preference was given to those studies that quantified endos ulfan exposure via analytical measurements and where appropriate, used a flow-through exposure regime. In some cases, however, these data were lacking (e.g., aquatic nonvascular plants), and toxicity values based on nominal concentrations were used to fulfill a data gap. In general, studies from the open literature do not provi de sufficient documentation for evaluating all or most of the OPPTS Guideline evaluation criteria. Consistent with the Overview Document (USEPA 2004), if sufficient docum entation was provided to judge that the open literature studies were scientifically valid and acceptable endpoints were determined that were below the toxicity reference values, they were typically classified as supplemental--but acceptable for quantitative use. If sufficient documentation was not provided to comprehensively judge the scien tific validity of the study but no information existed that suggested the study would be invalid, these studies were typically classified as supplemental--but acceptable for qualitative use. This classification was often applied to studies with nominal e xposure concentrations.

I. Acute Toxicity to Freshwater Fish

Submitted Data for Freshwater Fish Acute Values

Two freshwater fish toxicity st udies using the TGAI are required to establish the toxicity of endosulfan to fish. The preferred test spec ies are rainbow trout (a coldwater fish) and bluegill sunfish (a warmwater fish). Rainbow trout were the most sensitive (LC50 = 0.83
μ g/L) species tested (Table E-1). Sinc e the LC50 value was less than 0.1 mg/L, endosulfan is categorized as very highly toxi c to freshwater fish on an acute exposure basis. The freshwater fish acute toxicity test ing requirement (Guideline 72-1) is fulfilled (MRID 38806, 40094602, 40098001, 136999, 05008271).

Endosulfan Sulfate Toxicity

Acute toxicity of endosulfan sulfate on freshw ater fish was tested using bluegill sunfish. The 96-hour LC50 of endosulfan sulfate on bluegill was 3.8 μ g/L and is considered very highly toxic (MRID 46382604). Acute toxicity testing of endosulfan sulfate (99.9% radiopurity) was also tested with common carp (Gries and van der Kolk, 2000a; MRID

E-1

### 45421402), which also indicated that the degradat e is very highly toxic (96-hr LC50 = 2.2
μ g/L) to freshwater fish. The study was classified as s upplemental since it failed to comply with EPA’s recommended species and testing conditions. These results are consistent with studies of parent endosulf an where 96-hr LC50 values ranged from 0.37 to 3.3 μ g/L.

Formulated End Use Product

The acute toxicity of endosulfan to freshw ater fish was tested using technical end product. There was considerable variability fo r LC50 values within formulations tested; toxicity estimates ranged from 0.47 μ g/L to 5.6 μ g/L for the 50 WP and 33.7% formulations, respectively (Table E-2). Simila r to the results on technical grade active ingredient, rainbow trout were the most se nsitive species tested using technical end product. Of the formulated products tested, the 4% formulation was the least toxic with an LC50 estimate of 28 μ g/L. Based on the test results, the formulation of endosulfan is classified as being very highly toxic to freshwater fish species. The freshwater fish acute toxicity testing requirement using technical end product (Guideline 72-1) is fulfilled (MRID 00128655, BA007902, 250401A).

Additional Open Literature Information (ECO TOX) for Freshwater Fish Acute Values

All of the open literature acute values from EPA’s ECOTOX database for freshwater fish are presented in Figure E-1. The vertical line represents the toxicity reference value from the registrant’s data (0.83 ug/L). These data are primarily for an exposure duration of 4 days; however, other durations were included (e.g., 1 or 2 day expos ures) if there was no 4 day LC50 for a particular test. Tables E-3 and E-4 contains the acute LC50 values that are less than this toxicity reference valu e. When LC50 values for multiple exposure durations from the same test were available, only the value for 4 day exposure was included. There were 150 LC50 values from 35 open literature studies representing 31 freshwater fish species. Forty-five of these va lues were below the freshwater fish toxicity reference value (0.83 μ g/L) that was based on the inform ation provided by the registrant. These data are presented in nine differe nt studies and repr esent 13 species.

E-2Endosulfan: Freshwater Fish LC50s n = 150

0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0

0.1 1 10 100 1000 10000 Concentration (ug/L)Probability Rank

Figure E-1. Plot of the freshwat er fish endosulfan acute data.



### Forty-eight 96 hr LC50 values ar e available from an interla boratory comparison of acute tests for large subadults of the fathead minnow Pimephales promelas and fingerlings of the rainbow trout Oncorhynchus mykiss (Lemke1981—ECOTOX reference # 9479). All of the laboratories used the same technical grade endosulfan. For each fish species there are 24 LC50 values, 12 from static tests and 12 from flow-through tests. The static test values for fathead minnows ranged from 0.97 to 3.45 μ g/L (all values are above the toxicity reference value). The 12 flow-thr ough tests for this species had LC50s ranging from 0.29 to 1.91 μ g/L (seven of these values were be low the toxicity reference value). The static tests for rainbow trout ranged from 0.49 to 2.43 μ g/L (six of these 12 values were below the toxicity reference value) a nd the flow-through tests had values ranging from 0.17 to 0.75 μ g/L (all 12 values below the t oxicity reference value).

And additional set of fourteen 96 hr LC50s are presented in Sunderam et al. (1992— ECOTOX reference # 5850). These data are for ei ght species of fish from Australia. Six of the eight species had at le ast one LC50 values less than the toxicity reference value. These are the golden perch Macquaria ambigua , the common carp Cyprinus carpio , the eastern rainbow fish Melanotaenia duboulayi, the bony bream Nematolosa erebi , the harlequinfish Rasbora sp. and the rainbow trout Oncorhynchus mykiss. The endosulfan used was technical grade, and the common carp was the most sensitive species in this study and was the most sensitive freshwat er fish overall with an LC50 of 0.1 μ g/L. Seven tests in 1988 and 1989 used the striped bass Morone saxatilis and yielded similar LC50 values, ranging from >0.22 to 0.43 μ g/L (Fujimura et al. 1991—ECOTOX reference # 15472). All of the test s used larval fish, ranging in age from 6 to 45 days post hatch, and were conducted as flow-through tests.

Adult Asian swamp eels, Monopterus albus, were tested using a commercial endosulfan end product (33% purity—mixture of α and β endosulfans). The 96 hr LC50 obtained in this study was 0.42 μ g endosulfan/L (Hii et al. 2007—ECOTOX reference # 100663).

E-3

### Another study using males and females of Jenynsia multidentata —also with a commercial preparation (35% purity)—demonstr ated that males may be more sensitive than females; the 96 hr LC50s were 0.719 and 1.317 μ g endosulfan/L, respectively (Ballesteros et al. 2007—E COTOX reference # 103207).

Three different studies compared various forms of endosulfan; α and β endosulfans, technical grade mixture of the two, as well as commercial sources of powders and emulsified concentrates (Table E-5). In each case the α -endosulfan was the most toxic form, and the only form with an LC50 less than the toxicity reference value. Juveniles of Labeo rohita had an LC50 of 0.33 μ g/L for the α form; compared with 7.1 for the β

endosulfan and 1.0 to 1.25 for the technical grad e and commercial preparations (Rao et al. 1980—ECOTOX reference # 468). The snake-head catfish, Channa punctata had an LC50 of 0.16 μ g/L for α -endosulfan, and the other form s of endosulfan all had LC50s from 2.5 to 16 μ g/L (Devi, et al. 1981—ECOTOX refe rence # 15157). Finally, the 96 hr LC50 for α -endosulfan for the rainbow trout Oncorhynchus mykiss (Wan et al. 2005— ECOTOX reference # 87973) is 0.5 μ g/L, in addition, a 60/40 mixture of α - and β

endosulfans resulted in an LC50 of 0.7 μ g/L. The other forms of endosulfan had LC50s greater than the toxicity reference value. This included the only freshwater fish acute test from the ECOTOX data set that us ed endosulfan sulfate (LC50 of 1.4 μ g/L). In almost all cases the commercial end produc t was less toxic (based on ac tive ingredient ) than the technical grade.

Table E-1. Summary of freshwater fish acut e toxicity data submitted by registrant for endosulfanSpecies/Flowthrough or Static % ai 96-hour LC50 ( μ g/L) Toxicity Category MRID No. Author/Year Study Classification
Bluegill sunfish (Lepomis macrochirus) 96 2.08 very highly toxic BA007903/ EPA 1976 supplemental

Bluegill sunfish (Lepomis macrochirus) 100 1.7 very highly toxic 38806/ Buccafusco 1976 acceptable

Bluegill sunfish (Lepomis macrochirus) 96.6 3.3 very highly toxic 05014941/Pickering &Henderson 1966 supplemental

Bluegill sunfish (Lepomis macrochirus) static 96 1.2 very highly toxic 40094602/ Johnson&Finley 1980 supplemental

Rainbow trout (Oncorhynchus mykiss) static tech a 1.5 b very highly toxic 05003107/ Macek et al. 1969 supplemental

Rainbow trout (Oncorhynchus 96 1.1 b very highly toxic 40098001 Mayer & Ellerisieck supplemental

aThiodan : tested as μ g active ingredient per liter.
b Tested at several temperatures, this is the LC50 for the highes t temperature tested.


### E-4Species/Flowthrough or Static % ai 96-hour LC50 ( μ g/L) Toxicity Category MRID No. Author/Year Study Classification mykiss) static 1986

Rainbow trout (Oncorhynchus mykiss) static 95.9 0.83 very highly toxic 136999/ Fischer 1983 acceptable

Rainbow trout (Oncorhynchus mykiss) static 96 0.37 very highly toxic BA007902/ EPA 1976 supplemental

Channel catfish ( Ictalurus nebulosus ) static 96 1.5 very highly toxic 40094602/ Johnson&Finley 1980 supplemental

Fathead minnow (Pimephales promelas ) static 96 1.5 Very highly toxic 40094602/ Johnson&Finley 1980 supplemental

Fathead minnow (Pimephales promelas ) Flowthrough 99 0.86 c very highly toxic 05008271/ Macek et al. 1976 acceptable



### Table E-2. Summary of freshwater fish acut e toxicity data submitted by registrant for commercial endosulfan end products

Species/ Flow-through or Static % ai 96-hour LC50 ( μ g/L) Toxicity Category MRID No. Author/Year Study Classification

Bluegill sunfish (Lepomis macrochirus) 33.7 5.6 very highly toxic 00128656/ Kinter& Forbis 1983 supplemental

Bluegill sunfish (Lepomis macrochirus) 50WP 3.9 very highly toxic 00128655/ Kinter& Forbis 1983 acceptable

Rainbow trout (Oncorhynchus mykiss) static 50WP 0.47 very highly toxic BA007902/EPA 1976 acceptable

Rainbow trout (Oncorhynchus mykiss) static 50WP 2.3 very highly toxic 250401A acceptable

Rainbow trout (Oncorhynchus mykiss) static 33.7 2.7 very highly toxic 00128655/Kinter & Forbis 1983 supplemental

Rainbow trout (Oncorhynchus mykiss) static 4 28 very highly toxic 0033020/ Ludeman 1972 supplemental

Common carp (Cyprinus carpio) static 35 0.92 a very highly toxic 05004792/Basak & Konar 1976 supplemental

c 7-day LC50; however, similar to expected 96 hr LC50 (see text).


### E-5Species/ Flow-through or Static % ai 96-hour LC50 ( μ g/L) Toxicity Category MRID No. Author/Year Study Classification

Indian catfish (Heteropneustes fossilis) static 35 1.57 a very highly toxic 05004792/Basak & Konar 1976 supplemental

Mozambique tilapia (Tilapia mossambica) static 35 1.4 a very highly toxic 05004792/Basak & Konar 1976 supplemental

a 7-day LC50



### Table E-3. Summary of freshwater fish ac ute data from EPA’s ECOTOX database that was less than the toxi city reference value of 0.93 μ g/L. The shaded row is the lowest value.

Chemical Name Genus Species Common Name Dur (da) LC50 (ug/L) % a.i. Ref # Comment

α Endosulfan Channa punctata Snake-head catfish 4 0.16 100 15157 supplemental

Endosulfan Cyprinus carpio common carp 4 0.1 96.2 5850 Supplemental

α Endosulfan Labeo rohita Rohu 4 0.33 100 468 Supplemental

Endosulfan Macquaria ambigua Yellowbelly, Golden Perch 4 0.3 96.2 5850 Supplemental

Endosulfan Macquaria ambigua Yellowbelly, Golden Perch 4 0.5 96.2 5850 Supplemental

Endosulfan Melanotaenia duboulayi Eastern rainbow fish 4 0.5 96.2 5850 Supplemental

Endosulfan Morone saxatilis Striped bass 4 > 0.22 100 15472 Supplemental

Endosulfan Morone saxatilis Striped bass 4 0.23 100 15472 Supplemental

Endosulfan Morone saxatilis Striped bass 4 0.23 100 15472 Supplemental

Endosulfan Morone saxatilis Striped bass 4 0.29 100 15472 Supplemental

Endosulfan Morone saxatilis Striped bass 4 <0.33 100 15472 Supplemental

Endosulfan Morone saxatilis Striped bass 4 0.35 100 15472 Supplemental

Endosulfan Morone saxatilis Striped bass 4 0.43 100 15472 Supplemental

Endosulfan Nematolosa erebi Bony bream 4 0.2 96.2 5850 Supplemental

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.17 100 9479 a supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.24 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.26 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.26 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.27 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.29 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.3 100 9479 supplemental: measured



### E-6Chemical Name Genus Species Common Name Dur (da) LC50 (ug/L) % a.i. Ref # Comment

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.32 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.41 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.42 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.49 100 9479 supplemental: measured

α Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.4975 99.5 87973 supplemental

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.63 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.69 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.69 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.693 99 87973 supplemental

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.7 96.2 5850 supplemental

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.75 100 9479 supplemental

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.79 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.8 100 9479 supplemental: measured

Endosulfan Oncorhynchus mykiss Rainbow trout,donaldson trout 4 0.89 100 9479 supplemental: measured

Endosulfan Pimephales promel as Fathead minnow 4 0.29 100 9479 supplemental: measured

Endosulfan Pimephales promel as Fathead minnow 4 0.45 100 9479 supplemental: measured

Endosulfan Pimephales promel as Fathead minnow 4 0.73 100 9479 supplemental: measured

Endosulfan Pimephales promel as Fathead minnow 4 0.75 100 9479 supplemental: measured

Endosulfan Pimephales promel as Fathead minnow 4 0.76 100 9479 supplemental: measured

Endosulfan Pimephales promel as Fathead minnow 4 0.8 100 9479 supplemental: measured

Endosulfan Pimephales promel as Fathead minnow 4 0.81 100 9479 supplemental: measured

Endosulfan Rasbora sp. Harlequinfish, red rasbora 4 0.2 96.2 5850 supplemental

Endosulfan Tilapia nilotica Nile tilapia 2 0.69 100 20421 supplemental

a part of an EPA Interlaboratory Comparison Study—measured values and standard methods



### E-7

### Table E-4. Summary of freshwater fish acute da ta for commercial endosulfan end products from EPA’s ECOTOX database.

Chemical Name Genus Species Common Name Dur (da) LC50 (ug/L) % a.i. Ref # Comment

Endosulfan Jenynsia multidentata Onesi ded Livebearer 4 0.719 35 103207 supplemental

Endosulfan Monopterus albus Swamp or White Ricefield Eel 4 0.42 33 100663 supplemental



### Table E-5. Summary of freshwater fish acut e data from ECOTOX in which multiple sources of endosulfan were tested using th e same species. Note, most are not below the freshwater fish toxicity reference value.

Chemical Name Genus Species Common Name Dur (da) LC50 (ug/L) % a.i. Ref # Comment

α -Endosulfan Labeo rohita Rohu 4 0.33 - 468 supplemental
β− Endosulfan Labeo rohita Rohu 4 7.1 - 468 supplemental (α+β)

- Endosulfan Labeo rohita Rohu 4 1.1 96 468 supplemental

35% EC Labeo rohita Rohu 4 1 35 468 supplemental

4% Dust Labeo rohita Rohu 4 1.25 4 468 supplemental

α -Endosulfan Channa punctata Channel catfish 4 0.16 - 15157 supplemental

β− Endosulfan Channa punctata Channel catfish 4 6.6 - 15157 supplemental
(α+β)

- Endosulfan Channa punctata Channel catfish 4 4.8 96 15157 supplemental

35% EC Channa punctata Channel catfish 4 2.5 35 15157 supplemental

4% Dust Channa punctata Channel catfish 4 16 4 15157 supplemental

α -Endosulfan Oncorhynchus mykiss Rainbow trout 4 0.5 99.5 87973 supplemental

β− Endosulfan Oncorhynchus mykiss Rainbow trout 4 3.3 99.3 87973 supplemental
(α+β)

- Endosulfan Oncorhynchus mykiss Rainbow trout 4 0.7 99 87973 supplemental

40% EC Oncorhynchus mykiss Rainbow trout 4 4.6 40 87973 supplemental

50% WP Oncorhynchus mykiss Rainbow trout 4 3.5 50 87973 supplemental

Endosulfan Sulfate Oncorhynchus mykiss Rainbow trout 4 1.4 98 87973 supplemental



### E-8

### II. Chronic Toxicity to Freshwater Fish

Submitted Data for Freshwater Fish Chronic Values

An estimate of the chronic toxi city of technical grade endosul fan to freshwater fish was obtained in two test using life cycl e testing with the fathead minnows Pimephales promelas (Table E-6). In the study by Macek et al. (1976) there was no effect on survival or growth relative to the c ontrols in the F1 generation ( 40 weeks of exposure) or on egg production, hatchability, survival or growth in the F2 generati on (60 days of exposure) at 0.20 μ g/L. At the highest concentration tested (0.40 μ g/L) there also was no effect on the F1 generation after 30 or 60 days; howeve r, on day 170 there was a diluter malfunction and all of the fish died in this treatment. After 40 weeks, during the F2 exposure, eggs from the controls were added to the 0.40 trea tment and 1% of them hatched. Therefore, 0.40 μ g/L was the LOEC. In the second study (Dionne 2002), the estimated lowest observed effect concentration is 0.11 μ g/L based on reduced repr oduction (# eggs/female, # eggs/spawn, #spawns/female) and growth (reduced terminal length in F 0 

### males). The NOEC from this study is 0.056 μ g/L, and this value is the toxicity reference value against which the ECOTOX data are evaluated. The chronic toxicity testing requirement for freshwater fish (Guideline 72- 5) is fulfilled (MRIDs 45868601, 05008271).

Additional Open Literature Information (ECO TOX) for Freshwater Fish Chronic Values

There was only one study (Gormley and Teather 2003—ECOTOX Reference # 69838) which contained a chronic value less than the NOAEC value 0.056 μ g/L. The NOEC from that study for both growth and reproduction was 0.01 μ g/L; however, the next highest concentration tested was 0.1 μ g/L and the effects at this concentration were minimal (although statistically significant) to non-existent . The chronic value for the common carp was calculated usi ng the acute to chronic ratio for fathead minnow and the LC50 for carp. This resulti ng chronic value, 0.023 μ g/L (Table E-5), is used in the risk assessment for freshwater fish.

Table E-6. Summary of Freshwater flow-through fish life-cycle toxicity test using endosulfan
Species/Study Duration % ai NOEC/ LOEC

μ g/L Endpoints Affected MRID / Author Year Study Classification

Fathead minnow (Pimephales promelas) 340 days 99 NOEC = 0.2 LOEC= 0.4 Egg hatch 05008271/ Macek et al.1976 acceptable

Fathead minnow (Pimephales promelas) 99 NOEC = 0.056 LOEC= 0.11 Reduced reproduction and growth 45868601/ Dionne 2002 supplemental

Common carp (Cyprinus carpio) NOEC = 0.023 a -- -- --

aNOEC for common carp estimated using acute to chronic ratio for fathead minnow (0.86/0.20).



### E-9

### III. Acute Toxicity to Freshwater Invertebrates

Submitted Data for Freshwater Invertebrate Acute Values

A freshwater aquatic invertebrate toxicity testusing the TGAI is requ ired to establish the toxicity of endosulfan to aquatic invertebrates. The preferred test species is Daphnia
magna . Based on the available data (Table E-7) the estimated EC50 for technical grade endosulfan was 166 μ g/L using daphnia, 5.8 and 6 μ g/L for to species of Gammurus and 2.3 μ g/L for the stonefly Pteronarcys californica . Since the latter EC50 was less than 100
μ g/L, endosulfan is categorized as being very highly toxic to aquati c invertebrates on an acute exposure basis. The freshwater invert ebrate acute toxicity testing requirement (Guideline 72-2) is fulfilled (MRID 5008271, 40098001, 40094602).

Table E-7. Summary of freshwater invertebra te acute toxicity data submitted by registrant for endosulfan

Species/Duration % ai LC/EC50 ( μ g/L) Toxicity Category MRID No. Author/Year Study Classification Waterflea (Daphnia magna) 48 h 99 166 very highly toxic 5008271/ Macek et al. 1976 acceptable

Scud ( Gammurus faciatus) 96 h 96 6 very highly toxic 40098001/ Mayer & Ellersieck 1986 supplemental

Scud ( Gammurus lacustris ) 96 h 96 5.8 very highly toxic 40094602/ Johnson& Finley 1980 supplemental

Stonefly (Pteronarcys californica) 96 h 96 2.3 very highly toxic 40094602/ Johnson& Finley 1980 supplemental



### Acute toxicity testing endosulfan sulfate using Daphnia magna (99.4% active ingredient) indicated that the degradate is highly toxic (EC50 = 300 ug/L) to water fleas (Gries and van der Kolk, 2000b; MRID 45421403). The study wa s classified as supplemental since it was conducted using a water hardness (160 - 180 mg/L as CaCO 3 

### ) outside of the recommended range (40 - 49 mg/L as CaCO 3 

### ). These results are similar to the toxicity of the parent compound to aquatic invertebrates.

Additional Open Literature Information (ECO TOX) for Freshwater Invertebrate Acute Values

All of the open literature values from EPA’s ECOTOX database for freshwater invertebrates are presented in Table D-1 of Appendix D. A subset of this data representing the acute LC/EC50 values was co mpared against the toxicity reference value for freshwater invertebra te acute toxicity (2.3 μ g/L). These data are for exposure durations ranging from 0.5 to 4 days and are shown in Figure E-2. If values for multiple durations were available from a single test, th en only the longest dur ation value was used to make the comparison with the toxicity re ference value. There were 89 LC/EC50 values from 37 open literature studies representing 42 freshwater invertebrate species. Twentyone of these values were below the toxici ty reference value based on the information

E-10

### provided by the registrant. These data are presen ted in nine different studies and represent 13 species. These data are summarized in Table E-8. Note that the following discussion refers to some values that are not in Table E-8. This is because these data were not below the toxicity reference value.

Endosulfan: Freshwater Invertebrate LC50s n = 89

0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00

0.001 0.01 0.1 1 10 100 1000 10000 100000 LC50 (ug/L)Probability Rank

Figure E-2. Plot of the freshwater in vertebrate endosulfan acute data.


### One study conducted in Louisiana (USA) calcula ted the LC50 values for five different commercial pesticides, including Thiodan ( 33.7% endosulfan in xylene) using field collected microcrustaceans (Naqvi and Hawkins 1989—ECOTOX reference number 786). The LC50 values ranged from 0.1 to 0.9 μ g/L. The organisms were exposed as a community within which only four groups we re identified, and only to genus level — Diaptomus, Eucyclops, Alonella and Cypria . Because we cannot be certain that each of these genera represent only one species, and because we cannot be certain about the type and number of other organisms present in the exposure medium, these data were not considered in the risk assessment.

Two other studies with values below the toxici ty reference value, also were not used. One was eliminated because the author did not identify the source of endosulfan used (Magadza 1983—ECOTOX reference number 11147). Magadza presented LC50 values for four different invertebra tes, and the 48 h value for Daphnia longispina was 0.30 μ g/L. The other unused study (Pandian et al . 2000—ECOTOX reference number 93239) tested several pesticides, including Thiokill (35% endosulfan) using three different immature stages of the mosquito Culex quinquefasciatus . The LC50 values were 0.0021, 472 and 4509 μ g/L for the II instar larvae, IV instar larvae and pupae, respectively. The study was not used because the authors did not adequa tely describe the experimental design. The number of treatments was not given, and thei r methods only mention the fourth instar when describing the acute tests. However, they present results for three different immature stages.

E-11

### Leonard et al. (1999—ECTOX reference number 20012) exposed two mayfly nymphs, Jappa kutera and Atalophlebia australis , and larvae of a caddisfly, Cheumatopsyche sp. to technical grade endosulfan under static, nonrenewal conditions. Animals were exposed to endosulfan in the laboratory using wate r from the Namoi River, Australia. The concentrations of endosulfan were measured at the beginning of each test. Six values for the three species ranged from 0.4 to 1.8 μ g/L, the upper and lower values of this range were caddisfly data. However, the two values for Cheumatopsyche sp. were not used since the organism was not identified to specie s. The next lowest value from this study was for the mayfly Atalophlebia australis; however, this value (0.6 μ g/L) also is considered unacceptable. A later study (Hose et al. 2003; ECOTOX reference #73404) reported LC50 values an order of magn itude less sensitive for test using Atalophlebia spp. Correspondence with one of the coau thors from both studies (Ross V. Hyne, personal communication) clarifie d the difference in sensitivit y. Hyne indicated that the animals from the first study (Leonard et al. 1999) were likely stressed due to handling and transportation prior to the laboratory test s. This situation was corrected before the second study. Additional data are provided for Jappa kutera comparing its sensitivity to different forms of endosulfan (Leonard et al. 2001—ECOTOX re ference number 62479). These forms included α and β endosulfans, a technical grade (7:3 ratio of α and β ), and endosulfan sulfate. The two highest concentratio ns in the series used for each experiment were measured. The 96 hr LC50s were 0.7, 2.3, 1.8 and 1.2 μ g/L, respectively.

Two species of invertebrates were tested in beakers or cages held in flow-through artificial streams, the shrimp Paratya australiensis and the mayfly, Jappa kutera (Hose and Wilson 2005—ECOTOX reference numbe r 103213). The LC50s using technical grade endosulfan were 0.96 and 0.51 μ g/L for P. australiensis (two experiments), and 3.10 μ g/L for J. kutera. Although not stated in the paper, the authors refer to earlier work for the specific methods. That earlier work indicated that the e xposure concentrations were verified based on measured valu es. However, both LC50 values for P. australiensis are lower than the lowest measured test c oncentration—therefore th ese two tests can only be used qualitatively.

Three additional species of fr eshwater shrimp were tested in three separate studies. Juveniles of Palaemonetes paludosus were tested in a 96 hr renewal assay (renewed every 24 hr). The LC50 was 0.24 μ g/L; however, the concentrations were not measured and the source of the endosulfan was not reported (Kamthe 2002—ECOTOX reference number 93458). Adults of the tropical freshwater shrimp, Caridina laevis , were tested for acute toxicity using a variet y of commercial pesticides (Sucahyo et al. 2008—ECOTOX reference number 100785), including Thiodan 35EC (35% endosulfan). Exposures were static without renewal, and test results we re expressed based on c oncentrations of the active ingredient. The 96 hr LC50 for endosulfan was 1.02 μ g/L; however, the concentrations were not measured. The final LC50 value that was less than the toxicity reference value for freshwater invertebrates was for postlarvae of the freshwater prawn Macrobrachium rosenbergii (Lombardi, et al. 2001—ECOTOX reference number 61993). The exposure was to a commercial preparation (Thiodan CE); however, the nominal exposure concentrations were prep ared based on a calculated amount of active ingredient. The static, nonr enewal 96 hr LC50 was 0.93 μ g/L and the LC50 from a 96 hr renewal test (renewed every 24 hr) was 0.20 μ g/L. These results; however, are based on unmeasured concentrations.

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### Table E-8. Summary of freshwater invertebrate acute data from EPA’s ECOTOX database that was less than the toxicity reference value of 2.3 μ g/L. The shaded row is the value to be used for the risk assessment.

Chemical Name Genus Species Common Name Dur (da) LC50 (ug/L) % a.i. Ref # Comment

Endosulfan Alonella sp. Water flea 2 0.2 100 786 supplemental

Endosulfan Atalophlebia australis Mayfly 3 0.6 96 20012 invalid

Endosulfan Atalophlebia australis Mayfly 2 0.7 96 20012 invalid

Endosulfan Caridina laevis Smooth Caridina 4 1.02 100 100785 supplemental

Endosulfan Cheumatopsyche sp. Caddisfly 2 0.4 96 20012 invalid

Endosulfan Cheumatopsyche sp. Caddisfly 2 1.8 96 20012 invalid

Endosulfan Culex quinquefasciatus Southern house mosquito 1 0.0021 35 93239 supplemental

Endosulfan Cypria sp. Ostracod 2 0.9 100 786 supplemental

Endosulfan Daphnia longispina Water flea 2 0.3 100 11147 supplemental

Endosulfan Diaptomus sp. Calanoid copepod 2 0. 6 100 786 supplemental

Endosulfan Eucyclops sp. Cyclopoid copepod 2 0.1 100 786 supplemental

Endosulfan Jappa kutera Mayfly 4 0.7 100 62479 supplemental

Endosulfan Jappa kutera Mayfly 3 1 96 20012 invalid

Endosulfan Jappa kutera Mayfly 4 1.2 96 20012 invalid
Endosulfan sulphate Jappa kutera Mayfly 4 1.2 100 62479 supplemental

Endosulfan Jappa kutera Mayfly 4 1.8 100 62479 supplemental

Endosulfan Macrobrachium rosenbergii Giant river prawn 4 0.2 100 61993 supplemental
Endosulfan Macrobrachium rosenbergii Giant river prawn 4 0.93 100 61993 supplemental

Endosulfan Palaemonetes paludosus Riverine grass shrimp 4 0.24 100 93458 supplemental

Endosulfan Paratya australiensis Shrimp 2 0.51 96 103213 supplemental

Endosulfan Paratya australliensis Shrimp 2 0.96 96 103213 supplemental



### IV. Chronic Toxicity to Freshwater Invertebrates

Submitted Data for Freshwater Invertebrate Chronic Values

A freshwater aquatic invertebrate life-cycl e test using the TGAI is required for endosulfan since the end-use product is expect ed to be transported to water from the intended use site, and the acute EC50 is less than 1 mg/L. The preferred test species is Daphnia magna . The lowest observed effect concentration in daphnia ( Daphnia magna ) exposed to technical grade endosulfan was less than 7.0 μ g/L (Table E-9). The study examined the effects of endosulfan on thr ee consecutive generations of daphnids and noted that the poor survival of third genera tion in the control and the lowest treatment group (2.7 μ g/L) precluded drawing valid conclusi ons about the cumulative effects of exposure. However, the effects guidelines for a daphnid chronic to xicity test (OPPTS 850.1300) states that the durati on is 21 days. Therefore, we can evaluate the first generation exposure (22 days ) as a single test. The 2 nd

### and 3 rd

### generations in the original test were started by culling out 10 adu lt daphnids per replicate—making cumulative survival difficult to evaluate. It is note worthy that the NOEC estimate for daphnids (2.7

E-13

### μ g/L) is similar to the acute toxicity estimate for stoneflies (2.3 μ g/L) and does not provide a meaningful estimate of protection fo r freshwater invertebrates. However, the acute to chronic ratio for daphnids (LC50/NOEC: 166/ 2.7) could be used to predict the NOEC for the most sensitive species, the mayfly, Jappa kutera based on its LC50 (0.7 ug/L). The result of this calcula tion is a predicted NOEC of 0.011 μ g/L for mayflies. The chronic invertebrate toxicity testing requirement (Guidelin e 72-4) is fulfilled (MRID 5008271).

Table E-9. Freshwater aquatic invertebrate li fe-cycle toxicity test using endosulfan

Species % ai 21-day NOEC/LOEC ( μ g/L) Endpoints Affected MRID No. Author/Year Study Classification

Waterflea (Daphnia magna) 99 NOEC = 2.7 LOEC= 7.0 reduced survival 5008271/Macek et al. 1976 acceptable Mayfly (Jappa kutera -- NOEC = 0.011 a -- -- --

aPredicted based on acute LC50 for J. kutera (0.7 ug/L).divided by the acute to chronic ratio of 61.5 for daphnids (LC50/NOEC = 166/2.7).



### Sediment Toxicity

Endosulfan sulfate sediment toxicity tests were conducted for the midge, Chironomus
tentans , in two studies. The 20-day NOAEC and LOAEC determined from pore water were measured as 0.43 and 1.3 μ g/L, respectively, based on mortality (MRID 47318101). In a second study classified as suppleme ntal, the most sens itive NOAEC and LOAEC were determined as 2.7 and 3.8 μ g/L, respectively based on growth (MRID 46382605).

Additional Open Literature Information (ECO TOX) for Freshwater Invertebrate Chronic Values

There were only two papers from which grow th or reproduction effects values were potentially lower than the toxicity refere nce value for freshwater invertebrates (0.037
μ g/L). Both papers, however, were not us ed. The first paper (Pollino and Holdway 1999—ECOTOX reference number 20380) evaluated the acute and ch ronic effect of technical grade endosulfan on two species of hydra. The units for the chronic values presented in the authors’ Table 2 were a pparently mislabeled. The table shows NOEC values of 0.04 and 0.06 μ g/L; however, based on their Fi gure 2 and the text the units should have been mg/L. The actual NOEC values would therefore be 40 and 60 μ g/L, and not below the toxicity reference value.

The second paper evaluated the effect of endosulfan on the growth of juvenile Palaemonetes argentinus (Montagna and Collins 2007—ECOTOX reference number 101421). The test concentrations were not meas ured, and the size increase was similar for both the control and the treatment groups duri ng four molt cycles. The authors report that there is a significant relati onship between the cephalothorax length for the premolt and postmolt stages; however Table 2 is potential ly misleading and may give the erroneous

E-14

### idea that there were significant differences between the controls and the treatments, which there were not. V. Acute Toxicity to Estuarine/marine Fish

Submitted Data for Estuarine/marine Fish Acute Values

Acute toxicity testing with estuarine/mari ne fish using the TGAI is required for endosulfan because the end-use product is expe cted to reach this environment because of its use in coastal c ounties. The preferred test speci es is sheepshead minnow. Acute toxicity estimates for marine/estuarine fish ranged from 0.09 μ g/L to 0.38 μ g/L (Table E10); spot ( Leiostomus xanthurus ) was the most sensitive (LC50 = 0.09 μ g/L) species tested. Based on these data, endosulfan is ca tegorized as very highly toxic to marine/estuarine fish following an acute expos ure. The acute toxicity testing requirement using estuarine/marine fish (Guide line 72-3) is fulfilled (MRID 5005824).

Table E-10. Summary of estuarine/marine fi sh acute toxicity data submitted by registrant for endosulfan Species/Static or Flow-through % ai 96-hour LC 50 ( μ g/L) Toxicity Category MRID No./ Author Year Study Classification Striped Bass ( Morone saxatilis ) --- <1000 --- 05000819/Korn& Earnest 1974 supplemental Striped Bass ( Morone saxatilis ) 96 0.1 very highly toxic 00001328/Earnest 1970 supplemental Striped Mullet ( Mugil cephalis ) Flowthrough 96 0.38 very highly toxic 5005824/ Schimmel et al. 1977 acceptable

Pinfish (Lagodon rhomboides ) Flow-through 96 0.30 very highly toxic 5005824/ Schimmel et al. 1977 acceptable

Spot (Leiostomus xanthurus ) Flowthrough 96 0.09 very highly toxic 5005824/ Schimmel et al. 1977 acceptable



### Acute toxicity of endosulfan sulfate was test ed on the sheepshead minnow and resulted in a 96-hr LC50 of 3.1 μ g/L (MRID 46382603).

Additional Open Literature Information (E COTOX) for Estuarine/marine Fish Acute Values

All of the open literature values from EPA’ s ECOTOX database for estuarine/marine fish are presented in Table D-3 of Appendix D. There were no additional chronic values available from the ECOTOX database that we re less than the estuarine/marine acute toxicity reference value (0.09 μ g/L). However, there are LC50 values available for the preferred test species, the sheepshead minnow Cyprinodon variegatus. These data are from an interlaboratory comparison of acu te toxicity tests (Schimmel 1981—ECOTOX reference # 3740) and are displayed in Table E-11.

Table E-11. Summary of estuarine/mari ne acute data from EPA’s ECOTOX database for the fish ( Cyprinodon variegatus).

E-15Chemical Name Genus Species Common Name Dur (d) LC50 (ug/L) % a.i. Ref # Comment

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 0.34 100 3740 supplemental: measured

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 0.6 100 3740 supplemental: measured

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 0.83 100 3740 supplemental: measured

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 0.88 100 3740 supplemental: measured

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 1.1 100 3740 supplemental: measured

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 1.15 100 3740 supplemental: measured

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 1.2 100 3740 supplemental

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 1.4 100 3740 supplemental

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 2.7 100 3740 supplemental

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 2.81 100 3740 supplemental

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 2.87 100 3740 supplemental

Endosulfan Cyprinodon variegatus Sheepshead minnow 4 3.45 100 3740 supplemental



### VI. Chronic Toxicity to Estuarine/marine Fish

An estuarine/marine fish early life-stage toxi city test using the TGAI is required for endosulfan because the end-use product is expect ed to be transported to this environment from the intended use site and the aquatic acu te LC50 is less than 1 mg/L. The preferred test species is sheepshead minnow. No sheepshead minnow chronic data were submitted by the registrant; however, there are data fr om the ECOTOX database (Hansen and Cripe 1991—ECOTOX reference # 14143). These data are a summary of a 7-lab interlaboratory comparison of the ear ly life-stage toxicity test using Cyprinodon variegatus (Table E-12)

. All of the tests were flow-thr ough with measured concentration of endosulfan. The toxicant used wa s a technical grade mixture of 64% α -endosulfan and 36% β -endosulfan. The chronic toxi city data requirement for estuarine/marine fish (Guideline 72-4) is fulfille d. It is noteworthy that the NOEC estimate for sheepshead minnows (geomean = 0.36 μ g/L) is greater than the acute toxicity estimate for spot (0.09
μ g/L) and does not provide a meaningful estim ate of protection for estuarine/marine fish. However, the acute to chronic ratio fo r sheepshead minnows can be calculated by dividing the geometric mean of the acceptable LC50 values by the geometric mean of the chronic values (0.76/0.38). This ratio can be used to predict the NOEC for the most sensitive species, the spot, Leiostomus xanthurus. The result of this calculation is a predicted NOEC of 0.045 μ g/L for spot.

Table E-12. Estuarine/marine early life-stag e tests (28 da) for endosulfan (ECOTOX reference # 14143).

E-16Species NOEC (μg/L) LOEC (μg/L) Endpoint Study Classification Sheepshead minnow ( Cyprinodon variegates) 0.50 0.92 growth supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 0.31 0.54 growth supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 0.30 0.55 survival supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 0.29 0.47 growth supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 1.2 2.6 survival supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 0.21 0.41 survival supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 0.62 1.2 survival supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 0.58 1.2 survival supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 0.17 0.43 growth supplemental: measured Sheepshead minnow ( Cyprinodon variegates) 0.27 0.75 survival supplemental: measured Spot ( Leiostomus xanthurus ) 0.043 a
-- -- --

aPredicted based on acute to chronic ratio for sheepshead minnow (LC50/NOEC = 0.76/0.38)



### VII. Acute Toxicity to Estu arine/marine Invertebrates

Submitted Data for Estuarine/marine Invertebrate Acute Values Acute toxicity testing with estuarine/marine invertebrates using the TGAI is required for endosulfan because the end-use product is expe cted to reach this environment because of its use in coastal count ies. The preferred test species ar e mysid shrimp and eastern oyster. There was considerable variability in toxicityestimates for estuarine/marine invertebrates exposed to technical grade endosulfan. Estimated EC50 values ranged from 0.04 μ g/L to 790 μ g/L and represented a difference of four orders of magnitude (Table E-13). Based on the most sensitive EC50 (0.04 μ g/L) for pink shrimp, endosulfan is classified as very highly toxic to marine invertebrates. The estuarine/marine acute toxicity testing requirement (Guidelines 72-3b and 72 -3c) is fulfilled (MRID 128688, 5005824).

Table E-13 Summary of acute estuarine/marine invertebrate toxicity data submitted by the registrant for endosulfan

.

Species/Static or

Flow-through % ai. 96-hour EC 50 ( μ g/L)) Toxicity Category MRID No. Author/Year Study Classificatio n Eastern oyster (Crassostrea virginica) static embryo-larvae 96 0.45 very highly toxic 128688/Boeri& Ward/1983 supplemental

Fiddler crab (Uca pugnax) static 96 790 highly toxic 128688/Boeri& Ward 1983 supplemental Grass shrimp (Palaemonetes pugio) Flow-through 96 1.31 very highly toxic 5005824/ Schimmel et al. 1977 acceptable

Pink Shrimp 96 0. 04 very highly toxic 5005824/ acceptable



### E-17Species/Static or Flow-through % ai. 96-hour EC 50 ( μ g/L)) Toxicity Category MRID No. Author/Year Study Classificatio n (Penaeus duorarum) Flow-through Schimmel et al. 1977


### Acute toxicity of endosulfan sulfate was tested on the mysid shrimp and resulted in a 96- hr LC50 of 7.9 μ g/L, which is considered very highly toxic (MRID 46406401). Comparison among the mysid shrimp toxicity fo r endosulfan sulfate and the grass shrimp toxicity for the parent compound suggest the parent isomer is more toxic than the degradate.

Acute toxicity testing using t echnical end product (48% a.i.) on brown shrimp resulted in a 96-hr toxicity estimate of 0.24 μ g/L (Earnest 1970; Accession Number 00001328). Based on these data, technical end product is categorized as very highly toxic to estuarine/marine invertebrates.

Additional Open Literature Information (E COTOX) for Estuarine/marine Invertebrate Acute Values

In addition to the aforementioned acute mo rtality studies with estuarine/marine invertebrates, an additional study was identi fied in the open literature which reported several LC50 values for mysids ( Americamysis bahia; Schimmel 1981—ECOTOX reference # 3740). This study reported the resu lts of an interlaboratory comparison of acute toxicity tests using estuarine animals. There were five acute LC50 values based on measured, flow-through tests ranging from 0.38 to 1.29 μ g a.i./L (see Table E-14 ). While none of these values is lower than the LC50 for the pink shrimp listed above, their geometric mean (0.83 μ g a.i./L) will be used to establis h the acute to chronic toxicity ratio for estuarine/marine invertebrates used in the next section.

Table E-14. Summary of estuarine/mari ne acute data from EPA’s ECOTOX database for the mysid shrimp Americamysis bahia.
Chemical Name Genus Species Common Name Dur (d) LC50 (ug/L) % a.i. Ref # Comment

Endosulfan Americamysis bahia mysid shrimp 4 0.38 NR 3740 supplemental: measured

Endosulfan Americamysis bahia mysid shrimp 4 0.75 NR 3740 supplemental: measured

Endosulfan Americamysis bahia mysid shrimp 4 0.94 NR 3740 supplemental: measured

Endosulfan Americamysis bahia mysid shrimp 4 1.16 NR 3740 supplemental: measured

Endosulfan Americamysis bahia mysid shrimp 4 1.29 NR 3740 supplemental: measured Endosulfan Americamysis bahia mysid shrimp 4 0.24 NR 3740 supplemental Endosulfan Americamysis bahia mysid shrimp 4 0.46 NR 3740 supplemental Endosulfan Americamysis bahia mysid shrimp 4 0.73 NR 3740 supplemental Endosulfan Americamysis bahia mysid shrimp 4 1.12 NR 3740 supplemental Endosulfan Americamysis bahia mysid shrimp 4 1.47 NR 3740 supplemental



### E-18

### VIII. Chronic Toxicity to Estuarine/marine Invertebrates

An estuarine/marine invertebrate life-cycle t oxicity test using the TGAI is required for endosulfan because the end-use product may be applied directly to the estuarine/marine environment or is expected to be transporte d to this environment from the intended use site, and the following conditions are met: (1) th e pesticide is intended for use such that its presence in water is likely to be continuous or recurrent regardless of toxicity, (2) any aquatic acute LC50 or EC50 is less than 1 mg /L, (3) the EEC in water is equal to or greater than 0.01 of any acute LC50 or EC50 value, or, (4 ) the actual or estimated environmental concentration in water resul ting from use is less than 0.01 of any acute LC50 or EC50 value and any of the following co nditions exist: studies of other organisms indicate the reproductive physio logy of fish and/or invert ebrates may be affected, physicochemical properties indicate cumulative e ffects, or the pesticide is persistent in water ( e.g. , half-life greater than 4 days). No valid chronic data for estuarine/marine invertebrates exposed to technical endosulfan were submitted by the registrant. However, there are data from the ECOTOX data base (McKenney 1982—ECOTOX reference # 3736). These data are a summary of an interl aboratory comparison of the early life-stage toxicity test using Americamysis bahia— formerly Mysidopsis bahia (Table E-14)

. All of the tests were flow-through with measured c oncentration of technical endosulfan. The chronic toxicity data requirement for estuar ine/marine invertebrates (Guideline 72-4) is fulfilled. It is noteworthy that the NOEC estimate for mysid shrimp (geomean = 0.27
μ g/L) is greater than the acute toxi city estimate for pink shrimp (0.04 μ g/L) and does not provide a meaningful estimate of protection fo r estuarine/marine invertebrates. However, the acute to chronic ratio for mysids can be calculated by dividing the geometric mean of the acceptable LC50 values (Table E-15 by the geometric mean of the chronic values (0.83/0.27). This ratio can be used to predict the NOEC for th e most sensitive species, the pink shrimp, Penaeus duorarum. The result of this calcula tion is a predicted NOEC of 0.013 μ g/L for spot.

Table E-15. Estuarine/marine invertebrate chronic tests for endosulfan (ECOTOX reference # 3736).
Species NOEC ( μ g/L) LOEC ( μ g/L) Endpoint Affected Study Classification Mysid shrimp (Americamysis bahia) 0.33 0.71 survival/ reproduction supplemental: measured Mysid shrimp (Americamysis bahia) 0.52 1.12 survival supplemental: measured Mysid shrimp (Americamysis bahia) 0.14 0.31 survival supplemental: measured Mysid shrimp (Americamysis bahia) 0.28 0.60 survival supplemental: measured Mysid shrimp (Americamysis bahia) 0.22 0.28 survival supplemental: measured Pink shrimp ( Penaeus duorarum ) 0.013 a -- -- --

aPredicted based on acute to chronic ratio for mysid shrimp (LC50/NOEC = 0.83/0.27)



### E-19

### Chronic exposure to endosulfan sulfate was conducted for the mysid shrimp. The NOAEL and LOAEL were 0.38 and 0.73 μ g/L, respectively and the most sensitive endpoint was growth (MRID 46781601).

Endosulfan sulfate sediment toxicity test s were conducted for the estuarine amphipod, Leptocheirus plumulosus , in two studies (Table E-16 ). The 10-day NOAEC and LOAEC determined from pore water were measured as 27 and 45 μ g/L, respectively, based on mortality (MRID 46382606). In a 28-day study, the most sensitive NOAEC and LOAEC were determined as 1.58 and 4.0 μ g/L, respectively based on growth (MRID 46929001).

Table E-16. Toxicity of Endosulfan Sul fate to Aquatic Organisms Through Sediment Exposure.

Test Species Exposure Duration Endpoint (a) Pore Water ( μ g ai/L) Bulk Sediment (mg ai/kg dw) Study Classification (MRID)
Estuarine Amphipod, Leptocheirus plumulosus 10-d Survival: EC 50 LOAEC NOAEC 74 45 27 2.3 1.6 0.86 Supplemental (46382606)

Estuarine Amphipod, Leptocheirus plumulosus 28-d Growth: EC 50 LOAEC NOAEC >4.0 4.0 1.58 >1.2 1.2 0.48 Supplemental (46929001)

(a) information for the most sensitive endpoint is presented.



### IX. Toxicity to Aquatic-phase Amphibians

A number of studies involvi ng amphibian toxicity testing and endosulfan were identified in the open literature and are summarized be low. None of these studies provide reliable estimates of toxicity that may be used qua ntitatively in this risk assessment; however, they do provide some information regardi ng the hazard of endosulfan to amphibians. Although only supplemental information is av ailable for aquatic-phase amphibians, it appears that freshwater fish ar e substantially more sensitive.

Bernabò, et al. (2008—ECOTOX reference # 103223) and Brunelli, et al. (2009) evaluated the effect of endosulfan (99% purity) on ta dpoles of the common toad Bufo bufo. Four-day-old tadpoles were exposed for 96 hr in a static test. The 96 hr LC50 value based on nominal concentrations was 430 μ g/L (Bernabò, et al. (2008) . In a separate test, exposure to 200 μ g/L endosulfan resulted in an appare nt increase in mucus production by the gills, as well as inflammation of the gills . These changes occurred after only 24 hr and became more pronounced as the exposure continued (Bernabò, et al. 2008). The same group continued their work with endosulfan and this toad species looking for behavior and morphological differences (Brunelli, et al . 2009). Three concentrations and a control were used. There were no stat istically significant differences between the control and 10
μ g/L for survival, the rate of deformity or the number of toads metamorphosed (into Gosner stage 46—complete tail resorption). However, there was 77% and 97% mortality at 50 and 100 μ g/L, respectively. In addition, 100% of the surviving tadpoles exhibited various degrees of deformity and none of the survivors reach stage 46.

E-20

### Several papers evaluated the e ffect of endosulfan on tadpoles of Australian frogs, but all were unmeasured tests. Broomhall (2002—ECOTOX reference # 65732) exposed tadpoles of Litoria citropa to a single 0.8 μg/L nomina l concentration of endosulfan (Thiodan) under two different temperature regime s, one more variable than the other over the 96 hr test. At ambient temperatures (ca. 20 C) the survival was 89% in the endosulfan treatment (100% in the control). Under a sim ilar average temperature, but more variable, only 58% of the tadpoles survived, compared to 92% in the controls. After termination of the above test, all surv iving tadpoles were tran sferred to clean water and held at 19 C for 24 days. These tadpoles were then evaluate d for their ability to avoid predation by odonates. The endosulfan treated animals were more vulnerable to predation, measured by a significant decrease in the number of minutes elapsed until capture.

Broomhall and Shine (2003--ECOTOX refe rence # 71866) exposed tadpoles of the Australian tree frog Litoria freycineta to two different concentrations of endosulfan (source not given, but assume was same as Broomhall (2002)—Thiodan). There was no effect on survival after 96 hr exposure to 0.03 or 1.3 μ g/L a.i. relative to the controls. However, exposed tadpoles grew more slowly than controls. And the survivors were more vulnerable to odonate (dragon fly larvae) predations when tested 15 days after transfer to clean water. Broomall (2004—ECOTOX reference #73400) completed another series of experiments similar to the above, usi ng the same exposure concentrations with a different species, Limnodynastes peronii , with similar results.

Canadian frogs were evaluated based on unm easured concentrations. Green frog eggs and larvae ( Rana clamitans) were exposed to two diff erent scenarios using Thiodan 50WP (Harris, et al. 1998—ECOTOX refe rence #19300). In the first test series, Gosner stage 8 embryos through to stage 25 tadpoles were continuously exposed to a treatment for 13 days (two-thirds renewal of treatment solu tions every second day). The 96 hr LC50 was greater than the highest c oncentration tested (11,750 μ g/L a.i.); however, the 13 day LC50 was 15 μ g/L—suggesting that the earlier embryoni c stages were not as sensitive to endosulfan. Note, hatching occurred around da y eight of the test. A second set of exposures in this study consisted of the same exposure for the first 4 days, then a period of 7.5 days in reference water, followe d by a second 4-day exposure to the same concentrations. The 96 hr LC50 was again a greater than, and the 16 day LC50 was the same as that in the first test. The frequency of deformities at hatch also were calculated in the second test, resulting in an EC50 of 2,430 μ g/L.

The toxicity of Thiodan 4EC ( 40% emulsified concentrate) was evaluated (Berrill et al. 1998—ECOTOX reference # 19467) with newly ha tched and two-week-old tadpoles of the wood frog ( Rana sylvatica ), the American toad ( Bufo americanus ), and the green frog ( Rana clamitans ). Embryos of the wood frog were also tested, but were not as sensitive as tadpoles; therefore embryos of the other two species we re not tested. New hatched tadpoles of each species were exposed for 96 hr followed by a 10 day recovery/ observation period. Two-week of tadpoles were likewise exposed for 96 hr, and followed for and additional 7 day recovery period in clean water. Water concentrations of endosulfan were measured, but only after th e first hour of the exposure period and the treatments were apparently not renewed. There was essentially no mortality for either life stage for all three species during the firs t 96 hr; however, there was extensive post

E-21

### exposure mortality. Also, two-week old tadpole s were more sensitive than newly hatched tadpoles—no LC50s were calculated.

Early stage tadpoles of Bufo melaanostictus, Limnonectes limnocharis and Microphyla
ornata were exposed to a commercial end-use product of endosulfan (35EC) in 96-h acute toxicity tests (Dey and Gupta 2002—not in ECOTOX). Th e authors used 10 animals per container and three containers per treatment. They did not describe the chambers, the volume of water used, or the so urce of the water. Control mortality also was not described. The 96-h LC50s were base d on nominal concentrat ions and are 20, 13 and 0.16 μ g/L, respectively, for the three species listed above.

Finally, there is a preprint avai lable that evaluates the growth and survival of the northern leopard frog Rana pipiens tadpoles (Shenoy et al. in pres s). They exposed four-day-old tadpoles to Endosulfan 3EC (33% a.i.) for seve n weeks. Each individual was placed into a separate 1.5 L of exposure medium with th e water changed every 3 days. There were 15 individuals used for each trea tment. Besides the controls, treatments were 0.2, 1 and 5
μ g/L of the active ingredient. Concentrations were not measured. Gr owth rate (length) was not affected in surviving tadpoles in any treatment relative to the controls. No LC50 values were calculated; however, there was 100% mortality by day 12 and day 28 in the 5 and 1 μ g/L treatments, respectively. Based on the authors’ Figure 1 the control survival was approximately 80% at the end of the seven weeks and that in the 0.2 μ g/L treatment approximately 40%. Although no 96 hr data were presented, the figure suggests that there was little of no mortality during the first 4 days of the experiment.

X. Toxicity to Aquatic Plants There were no data on the toxicity of endosul fan to aquatic vascular or non-vascular plants submitted by the registrant. Plant data are generally not required unless the toxicant is an herbicide. There were several references available from the open literature: however, only one of these could be consid ered as following methods suitable to be considered acceptable (DeLorenzo et al 2002—ECOTOX reference # 65915). This study reported the 96 hr EC50 for the green alga Pseudokirchneriella subcapitatum (formerly Selenastrum capricoratum ) using a standard ASTM protocol . This species is one of the preferred species. The EC50 is 428 μ g/L based on nominal concen trations of technical grade endosulfan (98.6% a.i.). There were no da ta available for aquatic vascular plants.

References
The references that refer to an ECOTOX nu mber can be found in the reference list for Appendix D. The references listed below are for the data submitted to the Agency.

MRID Aquatic Citation References

Non-Guideline. Multi-Taxa Studies

40094602 Johnson, W.; Finley, M. (1980) Handbook of Acute Toxicity of Chemicals to Fish and Aquatic Invertebrates: Resource Publication 137. US Fish and Wildlife Service, Washington, D.C. 106 p



### E-2240098001 Mayer, F.; Ellersieck, M. (1986) Manual of Acute Toxicity: Inter- pretation and Data Base for 410 Chemicals and 66 Species of Freshwater An imals. US Fish & Wildlife Service, Resource Pub- lication 160. 579 p.

5008271 Macek, K.J.; Lindberg, M.A.; Sauter, S.; Buxton, K.S.; Costa, P.A. (1976 ) Toxicity of Four Pesticides to Water Fleas and Fathead Minnows. Duluth, Minn.: U.S. Environmental Protection Agency, Environmental Research Laboratory. (EPA report no. EPA-600/3-76-099; available from: NTIS, Springfield, VA; PB-262 912)



### Guideline 72-1. Acute Toxi city to Freshwater Fish

BA007903 U.S. EPA (1976) Report on the toxicity of thiodan to bluegill sunfish. U.S. EPA Chemical and Biological Investigations Branch, Beltsville, Maryland, static jar test No. 1023, unpublished report, 1976.

BA007902 U.S. EPA (1976) Report on the toxicity of thiodan to rainbow trout. U.S. EPA Chemical and Biological Investigations Branch, Beltsville, Maryland, static jar test No. 1033, unpublished report, 1976.

33020 Ludeman, J.A. (1972)Fish toxicity laboratory report, rainbow trout ( Salmo gairdnerii ) Test No. 496, I.D. Number 10033, USDA Reg. No. 70-126, Product name Kill-Ko Thiodan, Producer name, Rigo manufacturing Company, ARS-PR, ARC, Animal Biology Laboratory, Beltsville.

38806 Buccafusco, R.J. (1976) Acut e Toxicity of Endosulfan Technico to Bluegill ( Lepomis macrochirus ). (Unpublished study received Jul 28 , 1980 under 2749-487 ; prepared by EG&G, Bionomics, sub- mitted by Aceto Chemical Co., Inc., Flushing, N.Y.; CDL: 243084- A)

73120 Johnson, W.W. (1973) Letter sent to Robert E. Hawk dated Jul 12, 1973. Data for ethion and thiodan. (U.S. Fish and Wildlife Service, Fi sh-Pesticide Research Laboratory; unpublished study; CDL:120619-L)

101350 Ludemann, D.; Neumann, H. (1960) Versuc he uber die akute toxische wirkung neuzeitlicher kontaktinsektizide auf einsommerigc karpfen (Cyprinus carpio L.). Z. Angew Zool. 47(1):11- 33. (German text; also In unpublished subm ission received Nov 1, 1970 under unknown admin. no.; submitted by Hercules, Inc., Agricultural Chemicals, Wilmington, DE; CDL:005103-X)

109703 Oeser, H.; Gorbach, S.; Knauf, W. (1971) Endosulfane and the environment. Pages 17-22, In Giornate Fitopathologie, May, 1971, Udine, It aly. (S.l.:s.n.). (Also In unpublished submission received Nov 1, 1973 un der 3F1314; submitte d by FMC Corp., Middleport, NY; CDL:094299-L)

115694 U.S. Fish and Wildlife Serv ice, Fish-Pesticide Research Laboratory (19??) Toxicity of Ethion and Thiodan to Freshwater Fish. (Un- published study; CDL:248580-L)

128655 Kintner, D.; Forbis, A. (1983) Acute Toxicity of Thionex 3 EC to Rainbow Trout ...: Static Bioassay Report #29975. (Unpub lished study received May 31 , 1983 unde r 11678-25; prepared by Analyti- cal Bio-Chemistry Labo ratories, Inc., submitted by Makhteshim BeerSheva Chemical Works, Ltd., New York, NY; CDL:250401-A)

128656 Kintner, D.; Forbis, A. (1983) Acute Toxic ity of Thionex 3 EC to Bluegill Sunfish Static Bioassay Report #29974. (Unpub lished study received May 31 , 1983 unde r 11678-25;prepared by Analytical Bio-Chemistry Laboratories, Inc., submitted by Makh- teshim BeerSheva Chemical Works, Ltd., New York, NY; CDL: 250401-B)



### E-23136999 Fischer, R. (1983) The Effect of Hoe 002671 OI ZD96 0002 (Endosulfan, Active Ingredient 95.9%) on Salmo gairdneri (Rainbow Trout) in a Static Test: OEK83/019E; Document #A26006. (Unpublished study received Dec 27, 19 83 under 8340-13; prepared by Hoechst AG, W. Ger., submitted by American Hoeschst Corp., Somerville, NJ; CDL:252043-G)

148833 Turner, W.; Khalifa, S.; Casida, J. (1975) Toxaphene toxicant A. Mixture of 2,2,5-endo,6- exo,8,8,9,10-octachlorobornane and 2,2,5-endo,6-exo,8,9,9,10-octachlorobornane. J. Agric. Food Chem. 23(5):991-994.

151765 Kintner, D.; Forbis, A. (1983) Acute Toxicity of Thionex 50 WP to Rainbow Trout ( Salmo gairdneri ): Static Bioassay Report #29977. Unpublished report prepared by Analytical Biochemistry Labora- tories, Inc. 59 p.

151766 Kintner, D.; Forbis, A. (1983) Acute Toxicity of Thionex 50 WP to Bluegill Sunfish ( Lepomis macrochirus ): Static Bioassay Report # 29976. Unpublished report prepared by Analytical Bio-chemistry La boratories, Inc. 57 p.

250401A Kintner, D.; Forbis, A. (1983) Acute Toxicity of Thionex 50 WP to Rainbow Trout ( Salmo gairdneri ): Static Bioassay Report #29977. Unpublished report prepared by Analytical Biochemistry Laboratories, Inc. 59 p.

5003103 Amminikutty, C.K.; Rege, M.S. (1977) Effects of acute and chronic exposure to pesticides, Thiodan E_C_ 35 and Agallol '3' on the liver of widow tetra, Gymnocorymbus ternetzi (Boulenger). Indian Journal of Experimental Biology 15(3):197-200.

5003107 Macek, K.J.; Hutc hinson, C.; Cope, O.B. (1969) Th e effects of temperature on the susceptibility of bluegills and rainbow trout to selected pesticides. Bulletin of Environmental Contamination and Toxicology 4(3):174-183.

5003351 Reddy, T.G.; Gomathy, S. (1977) Toxicity and respiratory effects of pesticide, Thiodan on catfish, Mystus vittatus . Indian Journal of Environmental Health 19(4):360-363.

5004792 Basak, P.K.; Konar, S.K. (1976) Toxicity of six insecticides to fish. Geobios 3(6): 209-210.

5008271 Macek, K.J.; Lindberg, M.A.; Sauter, S.; Buxton, K.S.; Costa, P.A. (1976 ) Toxicity of Four Pesticides to Water Fleas and Fathead Minnows. Duluth, Minn.: U.S. Environmental Protection Agency, Environmental Research Laboratory. (EPA report no. EPA-600/3-76- 099; available from: NTIS, Springfield, VA; PB-262 912)

5014941 Pickering, Q.H.; Henderson, C. (1966) The acute toxicity of some pesticides to fish. Ohio Journal of Science 66(5):508-513.

45202909 Grande, M.; Andersen, S.; Berge, D. (1994) Effects of pesticides on fish: experimental and field studies. Norwegian Journal of Agri cultural Sciences Suppl. 13: 195-209.

45421402 Gries, T.; van der Kolk, J. (2000) (Carbon 14) Endosulfan sulfate (metabolite of (Carbon 14- Endosulfan) Acute Toxicity Test with Carp (Cyprinus carpio ) under Static Conditions: Lab Project Number: 1049.014.170: C010869. Unpublished study prepared by Springborn Laboratory (Europe) AG. 52 p.

45538301 Gries, T.; van der Kolk, J. (2000) (Carbon14)-Endosulfan Sulfate (Metabolite of (Carbon 14)-Endosulfan): Acute Toxicity Test with Rainbow Trout ( Oncorhynchus mykiss ) under Semi-Static Conditions: Lab Project Number: 1049.014.103: C016132. Unpublished study prepared by Springborn Laboratories AG. 54 p. {OPPTS 850.1075}

46382604 Cafarella, M. (2003) Endosulfan Sulf ate - Acute Toxicity to Bluegill Sunfish (Lepomis macrochirus ) Under Flow-Through Conditions. Project Number: 13798/6127, 33278/EBENX, B004613. Unpublished study prepared by Springborn Smithers Laboratories. 53 p. 

### Guideline 72-2. Acute Toxicity to Freshwater Invertebrates

E-24109704 Knauf, W.; Schulze, E. (1973) New Findings on the Toxicity of Endosulfan and Its Metabolites to Aquatic Organisms. (Unpublished study recei ved Nov 1, 1973 under 3F1314; prepared by Farbwerke Hoechst AG, W. Ger., submitted by FMC Corp., Middleport, NY; CDL:094299-M)

134394 Knauf, W.; Schulze, E. (1973) New Findings on the Toxicity of Endosulfan and Its Metabolites to Aquatic Organisms. (Unpublished study recei ved Nov 1, 1973 unde r unknown admin. no.; prepared by Farbwerke Hoechst AG, W. Ger., submitted by FMC Corp., Philadel- phia, PA; CDL:120346-J)

45421403 Gries, T.; van der Kolk, J. (2000) (Carbon 14) Endosulfan sulfate (metabolite of (Carbon 14- endosulfan) Acute Immobilisation Test with Daphnids ( Daphnia magna ) under Static Conditions: Lab Project Number: 1049.014.110: C012413. Unpublished study prepared by Springborn Laboratory (Europe) AG. 64 p.


### Guideline 72-3. Acute Toxicity to Estuarine/Marine Organisms

00001328 Earnest, R. (1970) Annual progress report: 1970 effects of pesticides on aquatic animals in the estuarine and marine environment. (Unpublished study received Nov 26, 1976 under 241-132; prepared by U.S. Fish and Wildlife Service, fi sh-Pesticide Research La boratory, submitted by American Cyanamid Co., Prin ceton, N.J.; CDL: 129448-W).

128658 Energy Resources Co., Inc. (1983) Acute Toxicity of Endosulfan to Embryos of the Eastern Oyster ( Crassostrea virginica ) and the Fiddler Crab ( Uca pugnax ). Final rept. (Unpublished study received May 31, 1983 under 8340-13; submitted by American Hoechst Corp., Somerville, NJ; CDL:250395-B)

128688 Boeri, R.H. and T.J. Ward. 1983. Acute toxicity of endosulfan to embryos of the eastern oyster (Crassostrea virginica) and the fiddler crab (U ca pugnax). Final Report. Submitted by American Hoechst Corp., Somerville, N.J.; prepared by En ergy Resources Co., Inc., Cambridge, Mass. Reg. No. 8340-13. Acc. No. 250395.

5000819 Korn S.; Earnest, R. (1974) Acute toxi city of twenty insecticides to striped bass Morone saxatilis . California Fish and Game, 60 (3); 128-131.

5003062 Roberts, D. (1975) The effect of pesticides on byssus formation in the common mussel, Mytilus edulis . Environmental Pollution 8(4):241-254.

5005824 Schimmel, S.C.; Patrick, J.M., Jr.; Wilson, A.J., Jr. (1977) Acute toxicity to and bioconcentration of endosulfan by estuarine animals. Pages 241-252, in Aquatic Toxicology and Hazard Evaluation. Edited by F.L. Mayer and J.L. Hamelink. Philadelphia, Pa.: American Society for Testing and Materials.

46382603 Cafarella, M. (2003) Endosulfan Sulfate - Acute Toxicity to Sheepshead Minnow ( Cyprinodon variegatus ) Under Flow-Through Conditions. Project Number: 13798/6138, EBENX014, B004571. Unpublished study prepared by Springborn Smithers Laboratories. 53 p.

46406401 Cafarella, M. (2003) Endosulfan Sulfate - Acute Toxicity to Mysids ( Americamysis bahia ) Under Flow-Through Conditions. Project Number: 13798/6137, EBENX015, B004570. Unpublished study prepared by Springborn Smithers Laboratories. 53 p.


### Guideline 72-4. Fish Early Life Stage/ Aquatic Invertebrate Life Cycle Study

3443 Reinert, H.K.; Parke, G.S.E. (1975) Report: Static 96 Hour Toxicity Study of Mobil Chemical Company Sample Modown 4 Lb./Gal. Flowable-M in Bluegill Sunfish and Rainbow Trout. (Unpublished study received Dec 12, 1975 under 2224-EX-10; prepared by Cannon Laboratories, Inc., submitted by Mobil Chemical Co., Industrial Chemic als, Richmond, Va.; CDL:223665-G)

29544 Pitcher, F. (1972) Kill-Ko Thiodan: T oxicity to Rainbow Trout: Test No. 496. (U.S. Agricultural Research Service, Pesticides Regulation Div., Animal Biology Laboratory,



### E-25unpublished report.)

33020 Pitcher, F. (1972) Kilt-Ko Thiodan: Toxicity to Rainbow Trout ( Salmo gairdneri ): Test No. 496. (U.S. Agricultural Re- search Service, Animal Biology Laboratory, unpublished study.)

33021 McCann, J.A. (1976) SA-50 Brand Thiodan 50 Wettable Powder: Toxicity to Rainbow Trout ( Salmo gairdneri ). (U.S. Environmental Protection Agency, Animal Biology Laboratory, unpublished study.)

33022 McCann, J.A. (1976) Metadan 2-2: Toxicity to Bluegill ( Lepomis macrochirus ). (U.S. Environmental Protection Agency, Animal Biology Laboratory, unpublished study.)

38306 Buccafusco, R.J. (1976) Acute Toxicity of Endo sulfan Technico to Bluegill ( Lepomis macrochirus ). (Unpublished study received Jul 28, 1980 under 2749-487; pr epared by EG&G, Bionomics, sub- mitted by Aceto Chemical Co., Inc., Flushing, N.Y.; CDL: 243084-A)

52151 McCann, J.A. (1976) Thiodan Tech 86%: Toxicity to Rainbow Trout ( Salmo garidneri ). (U.S. Environmental Protection Agency, Animal Biology Laboratory, unpublished study.)

52152 McCann, J.A. (1976) Thiodan Tech 86%: Toxicity to Bluegill (Lepomis macrochirus ). (U.S. Environmental Protection Agency, Animal Biology Laboratory, unpublished study.)

40462201 Graney, R. (1987) Progress Report No. 3: Endosulfan Pond Study. Unpublished study prepared by Battelle Columbus Laboratories. 79 p.

46510701 Heusel, R. (1991) Endosulfan - Substance, Technical (HOE 002671 00 ZD98 0005) Effect to Daphnia magna (Waterflea) In a 21-Day Reproduction Test (Method OECD). Project Number: CE90/076, A46561. Unpublished study prepared by Endosulfan Task Force. 246 p.

46781601 Cafarella, M. (2006) Endosulfan Sulfate - Life-Cycle Toxicity Test with Mysids ( Americamysis bahia ). Project Number: EBENM019, 13798/6178. Unpublished study prepared by Springborn Smithers Laboratories. 71 p. Relates to L0001048.



### Guideline 72-5. Life cycle fish

5003103 Amminikutty, C.K.; Rege, M.S. (1977) Effects of acute and chronic exposure to pesticides, Thiodan E_C_ 35 and Agallol '3' on the liver of widow tetra Gymnocorymbus ternetzi (Boulenger). Indian Journal of Experimental Biology 15(3):197-200.

45868601 Dionne, E. (2002) Endosulfan: The Chronic Toxicity to the Fathead Minnow ( Pimephales promelas ) During Full Life-Cycle Exposure: Lab Project Number: 13726.6140: B004189. Unpublished study prepared by Springborn Smithers Laboratories. 142 p.



### Guideline 850.1735 (Draft). Whole sedime nt acute freshwater invertebrates

46382605 Putt, A.E. (2004) Endosulfan Sulfate toxicity to midge ( Chironomus tentans ) during a 10-day sediment exposure. Springborn Smithers laboratories, Wareham, MA. Laboratory Report ID 13798.6142.

46382606 Putt, A. (2004) Endosulfan Sulfate - Toxicity to Marine Amphipods (Leptocheirus plumulosus) During a 10-Day Sediment Exposure. Project Number: 13798/6143, EBENX019, B004782. Unpublished study prepared by Springborn Smithers Laboratories. 83 p.

46929001 Putt, A. (2006) Endosulfan Sulfate - Toxicity to Estuarine Amphipods (Leptocheirus plumulosus) During a 28-Day Sediment Exposure. Project Number: 13798/6186, EBENX023. Unpublished study prepared by Springborn Smithers Laboratories. 60 p. Relates to L0001083.

47318101 Weltje, L; Pupp, A. (2008) Endosulfan sulfate – life cycle toxicity testing exposing midges ( Chironomus tentans ) to a test substance applied to sediment under static-renewal conditions following EPA test methods. Springborn Smithers Laboratories, Wareham, MA. Laboratory



### E-26Report ID. 13798.6185.



### E-27